环黄芪醇联合IL-7和IL-15对人脐带血T细胞体外扩增的影响  

Effects of cycloastragalus combined with IL-7 and IL-15 on the proliferation of human umbilical cord blood T cells in vitro

作　　者：郑武燕 冉文萍 家婷 赵日 曹倩 刘佳慧 邓彩云 郑雅文 陈巧 段曦 李华 ZHENG Wuyan;RAN Fengping;JIA Ting;ZHAO Ri;CAO Qian;LIU Jiahui;DENG Caiyun;ZHENC Yawen;CHEN Qiao;DUAN Xi;LI Hua(School of Basic Medicine,Chengdu University of Traditional Chinese Medicine Chengdu 610072,China;Department of Oncology,General Hospital of Western Theater Command,Chengdu 610083,China;Department of Gynecology and Obstetrics,First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China;Department of Basic Medicine,Southwest Jiao Tong University,Chengdu 610031,China;Department of Basic Medicine,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都中医药大学基础医学院,610072 [2]西部战区总医院肿瘤科,成都610083 [3]成都医学院第一附属医院妇产科,610500 [4]西南交通大学基础医学院,成都610031 [5]成都医学院基础医学院,610500

出　　处：《免疫学杂志》2019年第11期965-971,共7页Immunological Journal

基　　金：四川省科技厅应用基础重点项目(2018JY0440);四川省科技厅面上项目(19YYJC0242)

摘　　要：目的探究环黄芪醇(cycloastragenol,CAG)联合细胞因子IL-7和IL-15对人脐带血T细胞体外扩增和CD8^+T细胞亚群中干细胞样中心记忆性T细胞(stem cell-like central memory T cells,TSCM)比例的影响,为基于T细胞的肿瘤细胞免疫治疗研究奠定实验基础。方法采集健康产妇脐带血,通过密度梯度离心法分离获得脐带血单个核细胞(umbilical cord blood mononuclear cells,UCBMC),利用CD3/CD28抗体偶联磁珠刺激活化后,采用不同浓度的CAG(0、0.04、0.2、1、5μmol/L)联合IL-7和IL-15连续培养,利用多色流式细胞术分别测定T细胞的增殖程度以及CD8^+T细胞中TSCM比例的变化情况。结果 0.2μmol/L CAG与IL-7和IL-15细胞因子联用组促T细胞扩增作用显著高于其他浓度CAG与细胞因子IL-7和IL-15联用组或单独使用细胞因子IL-7和IL-15组。同时,0.2μmol/L CAG联用IL-7和IL-15组还可以促进CD8^+T中CD197^+CD45RA+细胞表型的表达,即维持CD8^+T中高比例的TSCM亚群。结论 0.2μmol/L CAG联合IL-7和IL-15可以显著提高活化T细胞扩增能力,并能保持CD8^+T细胞中高比例的TSCM亚群。This study was designed to explore the effect of cycloastragalus(CAG) combined with IL-7 and IL-15 on the proliferation of T cells, and the ratio of stem cell-like central memory T cells(TSCM). Human umbilical cord blood mononuclear cells(UCBMC) were obtained under informed consent based on guidance from healthy donors.Then, the UCBMC were isolated by Ficoll density gradient centrifugation, activated with anti-CD3/CD28 magnetic beads in RPMI-1640 with different concentration of CAG(0, 0.04, 0.2, 1, 5 μmol/L) combined with IL-7 and IL-15. The T cell expansion and the percentage of TSCMsubset were detected by flow cytometry. In the presence of IL-7,IL-15 and 0.2 μmol/L of CAG, the fold expansion of activated CD8^+T cells and the percentage of TSCMsubset were both significantly higher than those of other groups(P<0.05). In conclusion, 0.2 μmol/L of CAG combined with IL-7 and IL-15 can significantly improve the ability of T cell proliferation, and maintain a high ratio of TSCMsubgroup in CD8^+T cells.

关 键 词：环黄芪醇 T细胞 细胞扩增 干细胞样中心记忆性T细胞 

分 类 号：R730.5[医药卫生—肿瘤]